Illumina NovaSeq 6000 paired end sequencing; Single cell RNA-seq and TCR-seq of coding RNA from single cells in the mouse lung using 10x Chromium Single Cell 5' Reagent Kits User Guide (v2 Chemistry Dual Index)
Single cell RNA-seq and TCR-seq of coding RNA from single cells in the mouse lung using 10x Chromium Single Cell 5' Reagent Kits User Guide (v2 Chemistry Dual Index)
Briefly, lungs were harvested after perfusion with PBS. Lungs were first inflated with 0.1 mg/mL collagenase IV and DNase1 for 15 min at 37 °C. Single cell suspensions were prepared by mechanical dissociation of lung tissue through a 70-μm nylon mesh. Lung cells were suspended in PBS and layered on LSM Lymphocyte Separation (Medium MP Biomedical). Cells were centrifuged at room temperature for 20 min at 900 × g. Mononuclear cells were harvested from the gradient interphase. Followed by the manufacture's instructions for Chromium Next GEM Single Cell 5' Reagent Kits v2 (Dual Index) Followed by the manufacture's instructions for Chromium Next GEM Single Cell 5' Reagent Kits v2 (Dual Index)